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发酵微藻 P. lutheri 的细胞特性及其分离的活性肽对 MG-63 细胞成骨分化的影响。

Cellular properties of the fermented microalgae Pavlova lutheri and its isolated active peptide in osteoblastic differentiation of MG‑63 cells.

机构信息

Key Laboratory of Advanced Processing of Aquatic Products of Guangdong Higher Education Institution, College of Food Science and Technology, Guangdong Ocean University, Zhanjiang, Guangdong 524088, P.R. China.

School of Pharmacy, The University of Queensland, Brisbane, QLD 4072, Australia.

出版信息

Mol Med Rep. 2018 Jan;17(1):2044-2050. doi: 10.3892/mmr.2017.8087. Epub 2017 Nov 15.

Abstract

Fermented microalgae Pavlova lutheri (P. lutheri), the product of Hansenula polymorpha fermentation, exhibited an increase in alkaline phosphatase (ALP) activity in MG‑63 osteoblastic cells when compared to that of non‑fermented P. lutheri. Fractionation of the fermented P. lutheri resulted in identification of the active peptide [peptide of P. lutheri fermentation (PPLF)] with the sequence of EPQWFL. PPLF significantly increased ALP release from MG‑63 cells and mineralization in a dose‑dependent manner. In addition, the intracellular levels of ALP and osteocalcin (OCN) proteins were augmented by PPLF treatment. To identify the molecular mechanism underlying the effect of PPLF on osteoblastic differentiation, the phosphorylation levels of the mitogen‑activated protein kinases, p38, extracellular signal‑regulated kinases 1/2 and Jun, and nuclear factor (NF)‑κB were determined following PPLF treatment and the differences in expression were analyzed using p38 and NF‑κB selective inhibitors. These results concluded that PPLF from fermented P. lutheri induced osteoblastic differentiation by increasing ALP and OCN release in MG‑63 cells via the p38/p65 signaling pathway, indicating that PPLF supplement may be effective for therapeutic application in the field of bone health.

摘要

发酵微藻路德氏拟菱形藻(P. lutheri),经汉逊德巴利酵母发酵后,其碱性磷酸酶(ALP)活性在 MG-63 成骨细胞中较未发酵的 P. lutheri 有所增加。发酵后的路德氏拟菱形藻经分级分离后鉴定出活性肽[路德氏拟菱形藻发酵肽(PPLF)],其序列为 EPQWFL。PPLF 能显著增加 MG-63 细胞中 ALP 的释放,并呈剂量依赖性促进矿化。此外,PPLF 处理还能增加细胞内 ALP 和骨钙素(OCN)蛋白的水平。为了确定 PPLF 对成骨细胞分化的作用机制,用 PPLF 处理后测定了丝裂原活化蛋白激酶 p38、细胞外信号调节激酶 1/2 和 Jun 以及核因子(NF)-κB 的磷酸化水平,并使用 p38 和 NF-κB 选择性抑制剂分析了表达的差异。这些结果表明,发酵的路德氏拟菱形藻产生的 PPLF 通过增加 p38/p65 信号通路中 MG-63 细胞中 ALP 和 OCN 的释放来诱导成骨细胞分化,表明 PPLF 补充剂可能在骨骼健康领域的治疗应用中有效。

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